Department of Hematology, The Second Hospital of Shanxi Medical University, Taiyuan, Shanxi 030001, P.R. China.
Mol Med Rep. 2013 Feb;7(2):481-4. doi: 10.3892/mmr.2012.1194. Epub 2012 Nov 21.
The aim of this study was to investigate the mechanism of the apoptotic effect of brucine on human multiple myeloma (MM) cells. U266 cells (5x104) were plated in the presence or absence of brucine (0, 0.05, 0.1, 0.2 and 0.4 mg/ml) in 96‑well culture plates for 24‑72 h. The anti-proliferative response to brucine was assessed by MTT assay. Analysis of the cell cycle of U266 cells treated with or without brucine was performed using flow cytometry. The expression change of c-Jun following treatment with brucine or brucine plus the c-Jun N-terminal kinase (JNK)-specific inhibitor SP600125 was detected using RT-PCR. Brucine appeared to have an effect on apoptosis in a dose- and time‑dependent manner. Cell cycle analysis using flow cytometry revealed the accumulation of cells at the sub-G0/G1 phase. The apoptotic rates were 4.137, 10.55, 12.31, 27.67 and 29.67% (0, 0.05, 0.1, 0.2, 0.4 mg/ml brucine, respectively; P<0.01). The gray scale values were 0.7961 ± 0.007 and 0.4683 ± 0.003 (mRNA expression of c-Jun of U266 cells with or without SP600125, respectively). Concentrations of ≤ 0.4 mg/ml brucine induced apoptosis in U266 cells. Thus, brucine‑induced apoptosis in U266 cells occurs via the JNK signaling pathway and phosphorylation of c-Jun.
本研究旨在探讨马钱子碱诱导人多发性骨髓瘤（MM）细胞凋亡的机制。将 U266 细胞（5x104）接种于 96 孔培养板中，分别加入或不加入马钱子碱（0、0.05、0.1、0.2 和 0.4 mg/ml），培养 24-72 小时。采用 MTT 法检测马钱子碱对细胞增殖的抑制作用。采用流式细胞术分析 U266 细胞经马钱子碱或马钱子碱联合 c-Jun N-末端激酶（JNK）特异性抑制剂 SP600125 处理后的细胞周期变化。采用 RT-PCR 检测 U266 细胞经马钱子碱或马钱子碱联合 SP600125 处理后 c-Jun 的表达变化。结果显示，马钱子碱呈时间和剂量依赖性诱导 U266 细胞凋亡，细胞周期分析显示细胞在 sub-G0/G1 期积聚。凋亡率分别为 4.137%、10.55%、12.31%、27.67%和 29.67%（0、0.05、0.1、0.2、0.4 mg/ml 马钱子碱组，P<0.01）。灰度值分别为 0.7961±0.007 和 0.4683±0.003（加入或未加入 SP600125 时 U266 细胞 c-Jun 的 mRNA 表达）。浓度≤0.4 mg/ml 的马钱子碱诱导 U266 细胞发生凋亡。因此，马钱子碱诱导 U266 细胞凋亡是通过 JNK 信号通路和 c-Jun 的磷酸化实现的。
